LOCUS       sequence-351481-        3455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for generation of ClonMapper Recall Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3455)
  AUTHORS   Gardner A, Morgan D, Al'Khafaji A, Brock A
  TITLE     Functionalized Lineage Tracing for the Study and Manipulation of 
            Heterogeneous Cell Populations.
  JOURNAL   Methods Mol Biol. 2022;2394:109-131. doi: 
            10.1007/978-1-0716-1811-0_8.
  PUBMED    35094325
REFERENCE   2  (bases 1 to 3455)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2022"; volume: "2394:109-131. doi"; pages: " 
            10.1007/978-1-0716-1811-0_8"
COMMENT     Sequence Label: Recall-miniCMV-sfGFP
FEATURES             Location/Qualifiers
     source          1..3455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..131)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    177..232
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(231..250)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(640..1228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(720..739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1399..2190,2191..2259))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2022..2041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2260..2331)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2342..2566
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2555..2572
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2682..2720
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2717..2741
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2761..2763,2764..3455,1..22)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             join(2761..3455,1..19)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2791..2819)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3414..3434
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 catggatgag ctctacaaat aaaggtccgg ccgcactcct caggtgcagg ctgcctatca
       61 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
      121 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
      181 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
      241 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
      301 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
      361 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
      421 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
      481 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
      541 tcttctctta tggagatccc tcgacctgca ggtcaccaga agagctaacg gctcttccaa
      601 gcgggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1261 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1321 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1381 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1501 gggcttacca tctggcccca gtgctgcaat gataccgcgt gacccacgct caccggctcc
     1561 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1921 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2221 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2281 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagtg
     2341 gccatagagc ccaccgcatc cccagcatgc ctgctattga cttcccaatc ctcccccttg
     2401 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     2461 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     2521 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
     2581 cgggtttaaa ggagcgagac gaaggctcac tgtgcagact ggagcgcgtc tcctactatg
     2641 tcttcttact gaggatattg cctgaagctg gaagacgtaa tggtaggcgt gtacggtggg
     2701 aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga attcgccacc
     2761 atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     2821 gatgttaatg ggcacaaatt ttctgtcaga ggagagggtg aaggtgatgc tacaaacgga
     2881 aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     2941 gtcactactc tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg
     3001 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
     3061 aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
     3121 aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct cggacacaaa
     3181 ctcgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     3241 atcaaagcta acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac
     3301 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     3361 ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     3421 cttgagtttg taactgctgc tgggattaca catgg
//
